Table_1_Sinomenine Inhibits the Progression of Rheumatoid Arthritis by Regulating the Secretion of Inflammatory Cytokines and Monocyte/Macrophage Subsets.DOCX

Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory arthropathy associated with articular damage and attendant comorbidities. Even although RA treatment has advanced remarkably over the last decade, a significant proportion of patients still do not achieve sustained remission. The cause of RA is not yet known despite the many potential mechanisms proposed. It has been confirmed that RA is associated with dysregulated immune system and persistent inflammation. Therefore, management of inflammation is always the target of therapy. Sinomenine (SIN) is the prescription drug approved by the Chinese government for RA treatment. A previous study found that SIN was a robust anti-inflammation drug. In this study, we screened the different secretory cytokines using inflammation antibody arrays and qRT-PCR in both LPS-induced and SIN-treated RAW264.7 cells followed by evaluation of the ability of SIN to modulate cytokine secretion in a cell model, collagen-induced arthritis (CIA) mouse model, and RA patients. Several clinical indexes affecting the 28-joint disease activity score (DAS28) were determined before and after SIN treatment. Clinical indexes, inflammatory cytokine secretion, and DAS28 were compared among RA patients treated with either SIN or methotrexate (MTX). To explore the mechanism of SIN anti-inflammatory function, RA-associated monocyte/macrophage subsets were determined using flow cytometry in CIA mouse model and RA patients, both treated with SIN. The results demonstrated that SIN regulated IL-6, GM-CSF, IL-12 p40, IL-1α, TNF-α, IL-1β, KC (CXCL1), Eotaxin-2, IL-10, M-CSF, RANTES, and MCP-1 secretion in vivo and in vitro and reduced RA activity and DAS28 in a clinical setting. Furthermore, SIN attenuated CD11b+F4/80+CD64+ resident macrophages in the synovial tissue, CD11b+Ly6C+CD43+ macrophages in the spleen and draining lymph nodes of CIA mice. The percentage of CD14+CD16+ peripheral blood mononuclear cells was reduced by SIN in RA patients. These data indicated that SIN regulates the secretion of multiple inflammatory cytokines and monocyte/macrophage subsets, thereby suppressing RA progression. Therefore, along with MTX, SIN could be an alternative cost-effective anti-inflammatory agent for treating RA.

类风湿关节炎（Rheumatoid arthritis, RA）是一种慢性自身免疫性炎性关节病，可伴有关节损伤及相关并发症。尽管近十年来RA的治疗取得了显著进展，但仍有相当比例的患者无法达到持续缓解。虽然已提出多种潜在发病机制，但RA的具体病因至今尚未明确。已有研究证实，RA与免疫系统失调及持续性炎症密切相关，因此炎症调控始终是治疗的核心靶点。青藤碱（Sinomenine, SIN）是中国政府批准用于RA治疗的处方药。既往研究表明，青藤碱是一种强效抗炎药物。本研究首先通过炎症抗体芯片与qRT-PCR（quantitative real-time polymerase chain reaction）技术，在脂多糖（Lipopolysaccharide, LPS）诱导并经青藤碱处理的RAW264.7细胞中筛选差异分泌的细胞因子；随后分别在细胞模型、胶原诱导性关节炎（Collagen-induced arthritis, CIA）小鼠模型以及RA患者中，评估青藤碱对细胞因子分泌的调控作用。本研究检测了青藤碱治疗前后影响28关节疾病活动度评分（Disease Activity Score 28, DAS28）的多项临床指标，并对比了接受青藤碱与甲氨蝶呤（Methotrexate, MTX）治疗的RA患者的临床指标、炎性细胞因子分泌水平及DAS28评分。为探究青藤碱的抗炎作用机制，本研究通过流式细胞术，分别检测了青藤碱处理的CIA小鼠模型及RA患者体内与RA相关的单核细胞/巨噬细胞亚群。研究结果显示，青藤碱可在体内外调控IL-6、GM-CSF、IL-12 p40、IL-1α、TNF-α、IL-1β、KC（CXCL1）、Eotaxin-2、IL-10、M-CSF、RANTES及MCP-1的分泌，并在临床场景中降低RA疾病活动度与DAS28评分。此外，青藤碱可减轻CIA小鼠滑膜组织中CD11b+F4/80+CD64+驻留巨噬细胞，以及脾脏与引流淋巴结中CD11b+Ly6C+CD43+巨噬细胞的比例。在RA患者中，青藤碱可降低外周血单个核细胞中CD14+CD16+亚群的占比。上述数据表明，青藤碱可通过调控多种炎性细胞因子的分泌及单核细胞/巨噬细胞亚群比例，从而抑制RA的疾病进展。因此，青藤碱可与甲氨蝶呤联用，成为治疗RA的一种兼具成本效益的替代抗炎药物。